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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/Cell Meter™ Intracelluar NADH/NADPH Flow Cytometric Analysis Kit/15291/100 Tests
产品编号:15291
市  场 价:¥9900.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$495.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Cell Meter™ Intracelluar NADH/NADPH Flow Cytometric Analysis Kit/15291/100 Tests
商品介绍
Overview
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Ex/Em(nm)540/590
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellBIOLOGy
CellMetabolism
RelatedRedoxEnzymes
ThedetectionofintracellulardihydronicotinamideadeninedinucleotideNADHanditsphosphateesterNADPHisimportantfordiseasediagnosticsanddrugdiscovery.Ingeneral,theredoxcouplesNAD/NADHandNADP/NADPHplayacriticalroleinenergymetabolism,glycolysis,tricarboxylicacidcycleandmitochondrialrespiration.TheincreasedNAD(P)Hlevelincellsislinkedtotheabnormalproductionofreactiveoxygenspecies(ROS)andDNAdamage.However,duetothelackofsensitiveNAD(P)Hprobe,ithasbeenchallengingtodetectintracellularNAD(P)Hinbiologicalsystems.CellMeter™IntracellularNADH/NADPHFlowCytometricAnalysisKitprovidesanefficientmethodtomonitorintracellularNAD(P)Hlevelinlivecells.JZL1707NAD(P)HsensorhasbeendevelopedasanexcellentfluorescentprobefordetectingandimagingNADH/NADPHincells.TheprobebindNADH/NADPHtogeneratestrongfluorescencesignalwithhighsensitivityandspecificity.JZL1707NAD(P)HsensorcanbereADIlyloadedintolivecells,anditsfluorescencesignalcanbeconvenientlymonitoredusingflowcytometerinPEchannel.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Foreachsample,preparecellsin0.5mLserum-freemediumorbufferofyourchoiceatadensityof1×105to1×106cells/mL.
    Note1:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.Foradherentcells,gentlyliftthecellswith0.5mMEDTAtokeepthecellsintact,andwashthecellsoncewithserum-freemedia.
    Note2:JZL1707NAD(P)Hsensorisserumsensitive,thereforeit’srecommendedtokeepcellsinserum-freemediumorbufferofyourchoice.Alternatively,cellscanbepreparedandtreatedinregularfullmedium.Changetoserum-freemediumorbufferofyourchoicewhenincubationwithJZL1707NAD(P)Hsensor.

  2. Incubatecellswithtestcompoundsat37ºCforadesiredperiodoftimetostimulateintracellularNADH/NADPH.
    Note:Theappropriateincubationtimedependsontheindividualcelltypeandtestcompoundused.Optimizetheincubationtimeforeachexperiment.

  3. Add1μLofJZL1707NAD(P)Hsensor(ComponentA)into0.5mLcellsUSPension.Incubateat37ºCfor30-60minutes.
    Note:ForaNADH/NADPHpositivecontroltreatment:Jurkatcellswereincubatedwith100µMNADHorNADPHfor30minutesinserum-freemedium,andco-incubatedwithJZL1707NAD(P)Hsensorworkingsolutionat37ºCforanother30minutes.SeeFigure1fordetails.

  4. Washcellswithyourdesiredbufferonce.KeepcellsinAssayBuffer(ComponentB).

  5. MonitorthefluorescenceintensityatPEchannelusingaflowcytometer.Gateonthecellsofinterest,excludingdebris.
References&Citations
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1.         RubinTan,JianshaLi,XiaochunPeng,LipingZhu,LeiCai,TaoWang,YuanSu,KaikobadIrani,andQinghuaHu.GAPDHiscriticalforsuperiorefficacyoffemalebonemarrow-derivedmesenchymalstemcellsonpulmonaryhypertension.CardiovascRes,Oct2013;100:19-27.

 

2.         StephenY.Xue,ValeriaY.Hebert,DaniciaM.Hayes,CorieN.Robinson,MitziGlover,andTammyR.Dugas.NucleosideReverseTranscriptaseInhibitorsInduceaMitophagy-AssociatedEndothelialCytotoxicityThatIsReversedbyCoenzymeQ10Cotreatment.Toxicol.Sci.,Aug2013;134:323–334.

 

3.         KateJ.Roberts,AndrewCross,OlgaVasieva,RobertJ.Moots,andStevenW.Edwards.Inhibitionofpre-Bcellcolony-enhancingfactor(PBEF/NAMPT/visfatin)decreasestheABIlityofhumanneutrophilstogeneratereactiveoxidantsbutdoesnotimpairbacterialkilling.J.Leukoc.Biol.,Sep2013;94:481-492.

 

4.         WeijingCai,MayaRamdas,LiZhu,XueChen,GaryE.Striker,andHelenVlassara.Oraladvancedglycationendproducts(AGEs)promoteinsulinresistanceanddiabetesbydepletingtheantioxidantdefensesAGEreceptor-1andsirtuin1.PNAS,Sep2012;109:15888-15893.

 

5.         YueQiu,ClausTittiger,ClaudeWicker-Thomas,GaëlleLeGoff,SharonYoung,EricWajnberg,ThierryFricaux,NathalieTaquet,GaryJ.Blomquist,andRenéFeyereisen.Aninsect-specificP450oxidativedecarbonylaseforcuticularhydrocarbonbiosynthesis.PNAS,Sep2012;109:14858-14863.


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